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Pak. J. Bot., 44(5): 1497-1502, 2012. |
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Updated: 10-10-12 | ||||
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MOLECULAR CHARACTERIZATION AND PROKARYOTIC EXPRESSION OF ORF507 STERILITY-ASSOCIATED GENE IN CHILLI PEPPER (CAPSICUM ANNUM L.) CYTOPLASMIC MALE STERILITY
MING-HUA DENG1,2†, JIN-FEN WEN3†, JIN-LONG HUO4†, HAI-SHAN ZHU2, PEI WANG4, XIONG-ZE DAI1, ZHU-QING ZHANG1, HUI ZHOU2 AND XUE-XIAO ZOU1*
Abstract: 9704A is a cytoplasmic male sterility (CMS) of chilli pepper (Capsicum annuum L) that has been commercially used in chilli pepper hybrid production in China. However, little is known about the CMS-associated gene in chilli pepper 9704A.In this study, according to the sequence of relative references, a fragment of 593 bp including a 507-bp single-coding exon, Orf507 (GenBank accession No. GU357841) was cloned from leaves of chilli pepper CMS-9704A. Results revealed that the orf456-2 had one nucleotide deleted in codon 150 (nt450delC) and consists of a 507-bp single-coding exon which encodes 168 amino acid compared with the original sequence data of orf456. RT-PCR results showed that the Orf507 was transcribed in buds and leaves at all developmental stages of CMS-9704A and a hybrid F1, while no band was detected in the maintainer line. The nuclear restorer gene may restore fertility by post-transcriptional processing or translational/post-translational modification, but did not interfere with transcription of Orf507 directly. The entire coding region of the Orf507 was inserted into an expression vector pET32a(+) and transformed into Escherichia coli Rosetta. Expression of the fusion protein was successfully induced by isopropyl-β-D-thiogalactopyranoside. This study could establish a basis for understanding the molecular mechanism of chilli pepper CMS.
1Institute of Vegetable Crops, Hunan Academy of Agricultural Science, Changsha 410125, China 2College of Horticulture and Landscape, Yunnan Agricultural University, Kunming 650201, China 3Faculty of Modern Agricultural Engineering, Kunming University of Science and Technology, Kunming 650224, China 4Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China †Both these authors contributed equally to this work *Corresponding author’s: e-mail: zou_xuexiao@163.com; dengminghua1974@yahoo.com.cn |
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